Lab Bradford Assay and SDSPAGE Western Blot Analysis What i

Lab: Bradford Assay and SDS-PAGE & Western Blot Analysis

What is the purpose of the blocking reaction (step #1)? How does the blocking reagent accomplish this?

Solution

Bradford assay is an analytical process used for measurement of concentration of protein in a solution i.e. depends on the composition of amino acid of measured protein involving spectroscopy.   

SDS-PAGE -- Sodium Dodecyl polyacrylamide gel electrophoresis.This technique is used for protein separation by electrophoresis using polyacrylamide gel and SDS for protein denaturation.

Western blot analysis:-   It is an analytical technique used for protein detection in a sample of tissue homogenate or extract.   

The purpose of blocking reaction is for preventing nonspecific binding of antibodies otherwise they may bind to epitopes.It improves sensitivity of assay.These blocking reagents may involve salt solution and are with detergent or without detergent.Before transfer of samples they should be detected by antibodies and membrane should be kept for incubation and blocking reagents block unoccupied binding areas on membrane.